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Comparison of reagents for the determination of the functional activity of coagulation factors by the coagulation method.
Štorková, Marie ; Sadílek, Petr (advisor) ; Vrbacký, Filip (referee)
This bachelor thesis deals with the introduction of new more sensitive reagents for the determination of plasma coagulation factor activity of the intrinsic pathway into the hematology laboratory. The theoretical part deals with a brief description of the whole hemostasis process, individual coagulation factors, pathologies of hemostasis and determination of functional activity of coagulation factors. The practical part is devoted to a comparative study of the functional activity of coagulation factors VIII, IX, XI and XII. This study compares the results of the individual factors measured with the new STA C.K. Prest 5 reagents in combination with the respective immunodeficient plasmas with the results obtained by methods using the original STA PTT A reagents in combination with classical deficient plasmas. The measured values were then evaluated using the linear regression method and Bland-Altman plot to assess whether the methods using the new reagents could be implemented in the laboratory. The calculated values of the correlation coefficients of the individual methods are higher than r = 0.98, which together with the equations of the lines of the correlation plots indicate a close correlation between the results measured with the original reagents and those measured with the newly introduced...
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Analysis of body fluids on the automatic analyzer XT - 4000i
Tycová, Blanka ; Šatínský, Dalibor (advisor) ; Sadílek, Petr (referee)
ABSRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Analytical chemistry Candidate: Blanka Tycová Supervisor: Doc. RNDr. Dalibor Šatínský, Ph.D. Title of rigorous thesis: Analysis of body fluids on the automatic analyzer XT - 4000i This rigorous thesis focuses on the cytologic analysis of body fluids, which are analyzed in hematology laboratory OKH in the Pardubice regional hospital, a.s. These are the cerebrospinal fluid, pleural, pericardial and abdominal fluid, peritoneal dialysate and synovial fluid. From the measured data during the analysis of body fluids has been to focus on quality control and determine precision, truthfulness, linearity measurements, and the stability of peritoneal dialysate in the laboratory (21.7 řC) and cooler temperatures (6.9 řC). Statistical evaluation of the measured data showed that the analyzer provides the valid values. Graphic processing has been shown, that the measurement actually conducted linearly. The experiment confirmed, that greater stability shows a sample of peritoneal dialysate, that has been kept at a cooler temperature (6.9 řC). In this rigorous thesis were performed correlation analysis results of analysis the body fluids in the automatic analyzer XT - 4000i and microscopic method, which used a Fuchs - Rosenthal counting...
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Search for a suitable housekeeping gene for relative quantification in patients with the chronic lymphocytic leukemia
Ryšková, Barbora ; Vrbacký, Filip (advisor) ; Sadílek, Petr (referee)
5 ABSTRACT Charles University in Prague Faculty of Pharmacy in Hradec Králové Department of Biochemical Sciences Candidate: Bc. Barbora Ryšková Supervisor: Mgr. Filip Vrbacký Title of diploma thesis: Search for a suitable houskeeping gene for relative guantification in patients with the chronic lymfocytic leukemia Chronic lymphocytic leukemia (CLL) is the most common form of leukemia found in adults and that is why it is studied very intensively. The cause of CLL is still unknown and it is not known how to cure this lyphoproliferative disorder without high risk of complications that may occur with a bone marrow transplantation. Altered gene expression of angiogenic factors in CLL is still insufficiently known too, so we decided to study this topic in our laboratory. Our task was to establish method for quantification of proper housekeeping gene that can be used for normalization of results of quantification of expression of some angiogenic factors by real-time PCR. We decided to use Abl1 gene but that gene is alternatively spliced. Exon 1 containing variant is housekeeping only and we had to take it into account when choosing proper method. I managed to transform Escherichia coli cells and amplify plasmid containing Abl1 sequence. Plasmid was isolated, quantificated by spectrophotometer and used to...
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Comparison of basic coagulation methods - APTT, PT, FBG by mechanical and optical principle
Hricová, Júlia ; Fátorová, Ilona (advisor) ; Sadílek, Petr (referee)
In my thesis, the basic coagulation methods APTT, PT and fibrinogen were compared with the use of an optical and impedance principle. The coagulation analyser Sysmex CA 7000 was used for measuring coagulation tests using an optical method and the coagulation analyser AMAX Destiny Plus measured the formation of clots using an impedance method. In a theoretical part of my thesis, I described the process of formation of clots as well as the detection principles of the two analysers mentioned. I mainly focused on possible interferences which can influence the values of coagulation tests. I described how these interferences could develop, the mechanism of their influence on measured parameters and how their effects could be removed. In a practical part of my thesis, I described a preanalytic part of blood taking, the coagulation analysers and a principle of their measurement, an internal quality control and the reference values for these tests. For the coagulation tests APTT, PT and fibrinogen were used patient's normal, haemolytic, lipemic and icteric plasma. T-test done in Microsoft Excel was used to assess similarities or differences between the optical and measured impedance results. To see the differences between methods, graphs of mean values of each method were created. For the conclusion of my...
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Implementation of Antiplatelet Therapy Monitoring on the Analyzer Multiplate
Trenčanská, Šárka ; Fátorová, Ilona (advisor) ; Sadílek, Petr (referee)
This study describes the implementation of antiplatelet therapy monitoring on the analyzer Multiplate that works on the principle of impedance aggregometry. We used to determine four tests - ASPItest used for monitoring acetyl salicylic acid (ASA) therapy, ADPtest and ADPtestHS used to monitoring thienopyridines treatment and TRAPtest , which is used to monitoring treatment with GPIIb/IIIa inhibitors. The aim of this study was to verify the accuracy of this method for individual tests, to determine its own reference limits and compare them with literature data and on a group of cardiac patients receiving dual antiplatelet therapy (aspirin 100 mg per day and clopidogrel 75 mg per day) to assess any resistance to these drugs. Accuracy of the method we verified the group of 10 blood donors without any treatment. Each sample was repeatedly measured 5 times and from the measured values we calculated coefficients of variation. Our values of coefficients of variation from the values given by the manufacturer (CV 6 - 10 %) did not differ significantly (ASPItest: CV 8.7 %, ADPtest: CV 8.8 %, ADPtestHS: CV 13.3 % and TRAPtest: CV 12.1 %). We also verified on a group of 32 blood donors without any treatment reference ranges AUC for single tests, which were compared with reference limits from the literature....
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Possibilities of Laboratory Diagnostics of Lupus Anticoagulant Antibodies in the Special Hematology Laboratory of the Institute for Clinical and Experimental Medicine in Prague
Rybníčková, Soňa ; Fátorová, Ilona (advisor) ; Sadílek, Petr (referee)
Rybníčková Soňa THE POSSIBILITY OF ASSESSMENT OF LUPUS ANTICOAGULANT ANTIBODIES IN HEMATOLOGY LABORATORY IKEM IN PRAGUE Bachelor thesis Charles University in Prague, Faculty of Pharmacy in Hradec Králové Laboratory Technician Background Diagnosis of the Lupus Anticoagulant is very complicated due to a missing completely reliable screening test and whenever the presence of prolonged clotting times on a routine plasma test is detected, the other potential causes besides LA is needed to exclude. The aim of the thesis is focused on a possibilities of testing the Lupus Anticoagulant in our Clinical Haematology Department in the Institute for Clinical and Experimental Medicine inclusive of ambiguity measured results interpretation and a necessity to abide by the conditions of a preanalytic phase. In addition, clinical manifestation and laboratory criteria assessment as well as a possible interferences in Lupus Anticoagulant testing are also discussed. Methods The theoretical part is focused on a basic mechanism of haemostasis, on an effect of antiphospholipid antibodies, the diagnostic kriteria inclusive of a treatment guidelines. The practical part is focused on importance of the preanalytical phase, principally on necessity of a double centrifugation tube method (to obtain platelet pure plasma). More over is...
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